JP2020503879A - Lepidopteran昆虫に対する活性を有する殺虫毒性タンパク質 - Google Patents
Lepidopteran昆虫に対する活性を有する殺虫毒性タンパク質 Download PDFInfo
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Abstract
Description
本出願は、2017年1月12日出願の米国仮出願番号62/445,313の利益を主張するものであり、その全体は参照により本明細書に組み込まれる。
本明細書では、2018年1月11日に作成された「MONS434WO―sequence listing.txt」という名称のファイルを含有するコンピュータ可読形式の配列表を提供する。このファイルはMS−Windows(登録商標)による測定において166,134バイトであり、その全体は参照により本明細書に組み込まれる。この配列表は配列番号:1〜配列番号:12から構成される。
Bacillus thuringiensis(Bt)細菌種が産生する昆虫阻害性タンパク質は当技術分野において周知である。特定のBtタンパク質を使用し、1種または複数種のこのようなタンパク質を含有する農業的に許容される製剤を植物に散布して、殺虫剤として有効な量のこのようなタンパク質を含有するように配合した組成物で種子をコーティングすることにより、または、結果的に有効となる1種または複数種のタンパク質を植物/種子内で発現させることにより、作物植物の農業害虫を防除することができる。
配列番号:1は、BCW 001 Lepidopteran毒性タンパク質をコードする天然B.thuringiensis株EG4384のヌクレオチド配列である。
本実施例は、毒性タンパク質BCW 001の発見及び解析、ならびに、キメラ毒性BCW 002及びBCW 003の構築について教示する。
本実施例は、BCW 001、002及び003タンパク質の効果的なLepidopteran害虫防除生物活性について教示する。
本実施例は、BCW 001のドメインII及びIIIをその他のCry1毒素などの対応するドメインに置換する際に、このようなドメインが、クロヨトウムシ生物活性をその他のCry1毒性タンパク質に伝えるのに十分であることについて教示する。
本実施例では、様々なLepidopteran害虫に対するバイオアッセイで試験を行った際のBCW 001、002及び003の毒性特性について説明する。
本実施例は、植物、植物に使用する植物ベクター及び構築物の作製、ならびに、本発明のタンパク質を発現する植物の生成に使用する、本発明のタンパク質をコードする合成配列の構築について教示する。
Claims (16)
- (a)配列番号:2、配列番号:4、配列番号:6、配列番号:8、配列番号:10及び配列番号:12からなる群から選択される配列のアミノ酸1〜607を含むアミノ酸配列を有する殺虫性タンパク質またはその殺虫性フラグメント、
(b)(a)の前記アミノ酸配列に対して少なくとも95%の同一性を示すポリペプチドフラグメント、
(c)(a)の前記アミノ酸配列に対して少なくとも84%の同一性を示すポリペプチドフラグメント、
(d)(a)の前記アミノ酸配列に対して少なくとも64%の同一性を示すポリペプチドフラグメント、または、
(e)配列番号:2、4、6、8、10または12のいずれか1つに記載の7位から607位までの前記アミノ酸配列を有する殺虫性タンパク質、
をコードするヌクレオチド配列を含むポリヌクレオチド構築物であって、
前記ヌクレオチド配列は異種プロモーター配列に機能的に連結する、
前記ポリヌクレオチド構築物。 - (a)配列番号:2の256位から606位までに記載のアミノ酸配列と、
(b)配列番号:4の257位から607位までに記載のアミノ酸配列と、
(c)配列番号:6の257位から607位までに記載のアミノ酸配列と、
を含む、
クロヨトウムシLepidopteran種に対する毒性を有するタンパク質。 - 前記殺虫性タンパク質は、Spodoptera frugiperda、Spodoptera exigua、Spodoptera litura、Mamestra configurata、Striacosta albicosta、Trichoplusia ni、Pseudoplusia includens、Anticarsia gemmatalis、Hypena scabra、Heliothis virescens、Agrotis subterranea、Pseudaletia unipuncta、Agrotis ipsilon、Agrotis orthogonia、Ostrinia nubilalis、Amyelois transitella、Crambus caliginosellus、Herpetogramma licarsisalis、Homoeosoma electellum、Elasmopalpus lignosellus、Cydia pomonella、Endopiza viteana、Grapholita molesta、Suleima helianthana、Plutella xylostella、Pectinophora gossypiella、Lymantria dispar、Alabama argillacea、Archips argyrospila、Archips rosana、Chilo suppressalis、Cnaphalocrocis medinalis、Crambus caliginosellus、Crambus teterrellus、Diatraea grandiosella、Diatraea saccharalis、Earias insulana、Egrias vittella、Helicoverpa armigera、Helicoverpa zea、Heliothis virescens、Herpetogramma licarsisalis、Lobesia botrana、Pectinophora gossypiella、Phyllocnistis citrella、Pieris brassicae、Pieris rapae、Plutella xylostella、及びTuta absolutaからなる群から選択されるLepidopteran種に対する活性を示す、請求項1に記載のポリヌクレオチド構築物。
- Spodoptera frugiperda、Spodoptera exigua、Spodoptera litura、Mamestra configurata、Striacosta albicosta、Trichoplusia ni、Pseudoplusia includens、Anticarsia gemmatalis、Hypena scabra、Heliothis virescens、Agrotis subterranea、Pseudaletia unipuncta、Agrotis ipsilon、Agrotis orthogonia、Ostrinia nubilalis、Amyelois transitella、Crambus caliginosellus、Herpetogramma licarsisalis、Homoeosoma electellum、Elasmopalpus lignosellus、Cydia pomonella、Endopiza viteana、Grapholita molesta、Suleima helianthana、Plutella xylostella、Pectinophora gossypiella、Lymantria dispar、Alabama argillacea、Archips argyrospila、Archips rosana、Chilo suppressalis、Cnaphalocrocis medinalis、Crambus caliginosellus、Crambus teterrellus、Diatraea grandiosella、Diatraea saccharalis、Earias insulana、Egrias vittella、Helicoverpa armigera、Helicoverpa zea、Heliothis virescens、Herpetogramma licarsisalis、Lobesia botrana、Pectinophora gossypiella、Phyllocnistis citrella、Pieris brassicae、Pieris rapae、Plutella xylostella、及びTuta absolutaからなる群から選択されるLepidopteran種に対する生物活性を更に含む、請求項2に記載のタンパク質。
- 請求項1に記載のポリヌクレオチド構築物を含むベクター。
- 請求項1に記載のポリヌクレオチド構築物を含む宿主細胞であって、前記宿主細胞は、細菌細胞、イースト細胞及び植物細胞からなる群から選択される、前記宿主細胞。
- 前記植物細胞は、アルファルファ、バナナ、大麦、インゲン、ブロッコリー、キャベツ、アブラナ属、人参、キャッサバ、トウゴマ、カリフラワー、セロリ、ヒヨコマメ、ハクサイ、柑橘類、ココナッツ、コーヒー、トウモロコシ、クローバー、綿、ウリ科、キュウリ、ダグラスモミ、ナス、ユーカリ、亜麻、ニンニク、ブドウ、ホップ、リーキ、レタス、テーダマツ、キビ、メロン、ナッツ、オート麦、オリーブ、タマネギ、観賞植物、ヤシ、牧草、エンドウ、ピーナッツ、コショウ、キマメ、マツ、ジャガイモ、ポプラ、カボチャ、ラジアータマツ、ダイコン、菜種、イネ、根茎、ライ麦、ベニバナ、低木、モロコシ、ダイオウマツ、ダイズ、ホウレンソウ、トウナス、イチゴ、サトウダイコン、サトウキビ、ヒマワリ、スイートコーン、モミジバフウ、サツマイモ、スイッチグラス、茶、タバコ、トマト、ライ小麦、芝草、スイカ及び小麦の植物細胞からなる群から選択される、請求項6に記載の宿主細胞。
- 請求項1に記載のポリヌクレオチド構築物を含む植物。
- 請求項8に記載の植物が生成する種子であって、前記種子は検出可能量の前記ポリヌクレオチド構築物を含む、前記種子。
- 前記植物が生成する種子、花粉、子孫、植物細胞、植物組織及び日用品は、検出可能量の前記ポリヌクレオチド構築物を含む、請求項8に記載の植物。
- 検出可能量の請求項1に記載のポリヌクレオチド構築物を含む生物試料。
- Lepidopteran害虫種を防除するための殺虫剤として有効な量の請求項2に記載のタンパク質と、
(a)前記タンパク質とは異なり前記同一のLepidopteran種に対する毒性を同様に有する薬剤であって、前記薬剤は、前記タンパク質とは異なるアミノ酸配列を有するポリペプチド、RNA分子及び化合物からなる群から選択される、前記薬剤、または、
(b)Cry1A、Cry1Ab、Cry1Ac、Cry1A.105、Cry1Ae、Cry1B、Cry1C、Cry1D、Cry1E、Cry1F、Cry1G、Cry1H、Cry1I、Cry1J、Cry1K、Cry1L、Cry2A、Cry2Ab、Cry2Ae、Cry4B、Cry6、Cry7、Cry8、Cry9、Cry15、Cry43A、Cry43B、ET35、ET66、TIC400、TIC800、TIC807、TIC834、TIC853、TIC1415、VIP3A、VIP3Ab、AXMI殺虫性タンパク質、DIG殺虫性タンパク質、eHIP、及びVIPタンパク質からなる群から選択される薬剤、
を提供する組成物。 - 別の殺虫剤を更に含む請求項12に記載の組成物であって、前記別の薬剤は、Cry1C、Cry3A、Cry3B、Cry34、Cry35、Cry51Aa1、ET29、ET33、ET34、ET70、TIC407、TIC417、TIC431、TIC901、TIC1201、TIC3131、5307、DIG−10、Axmi184、Axmi205及びAxmiR1からなる群から選択される、前記組成物。
- 請求項1に記載のポリヌクレオチド配列を含む種子を生成するための方法であって、前記方法は、
(a)前記ポリヌクレオチドを含む1つまたは複数の種子を植えることと、
(b)前記種子から植物を栽培することと、
(c)前記植物から多数の種子を収穫することであって、前記種子は前記ポリヌクレオチド構築物を含む、前記収穫することと、
を含む、
前記方法。 - (a)DKB89614−9、MON801、MON802、MON809、MON810、MON863、MON88017、MON89034、イベント4114−3、イベント5307、DAS59122−7、Bt10、Bt11、Bt176、CBH−351、DKB−83614−9、MIR162、MIR604、TC1507、TC6275、イベント676、イベント678、イベント680、イベント98140、DAS40278−9、DKB89790−5、MON21−9、HCEM485、MON832、MON87427、NK603、T14、T25及びVCO01981−5からなる群から選択されるトウモロコシ植物形質転換イベントと、
(b)MON87751、DAS81419−2、MON87701、A2704−12、A2704−21、A5547−127、A5547−35、CV127、DAS44406−6、DAS68416−4、DP356043、FG72、MON4032、ACS−GM003−1、MON87705、MON87708、MON89788、W62、W98、及びGFM Cry1Aからなる群から選択されるダイズ植物形質転換イベントと、
(c)DAS24236−5、DAS21023−5、イベント31707、イベント31803、イベント31807、イベント31808、イベント42317、BNLA−601、COT102、COT67B、イベント1、GHB119、GK12、MON15985、MLS9124、MON1076、MON531、MON757、T303−3、T304−40、SGK321、イベント19−51a、GHB614、LLCotton25、MON88701、MON88702、MON1445、MON1698及びMON88913からなる群から選択される綿植物形質転換イベントと、
(d)サトウキビ植物形質転換イベントNXI−1Tと、
(e)LLRICE06、LLRICE601、LLRICE62、GM−A17054及びGM−A17054からなる群から選択されるイネ植物形質転換イベントと、
を更に含む、
請求項8に記載の植物。 - 機能的に連結する、
(a)Cry1AドメインIアミノ酸配列を含む第1のペプチドセグメントと、
(b)BCW 001ドメインII−ドメインIIIアミノ酸配列を含む第2のペプチドセグメントと、
を含むLepidopteran毒性タンパク質であって、
前記Lepidopteran毒性タンパク質は、クロヨトウムシ(A.ipsilon)、オオタバコガ(H.zea)、西部インゲンヨトウムシ(S.albicosta)、ヨーロッパアワノメイガ(O.nubilalis)、南西部アワノメイガ(D.grandiosella)、イラクサギンウワバ(T.ni)、ダイズシャクトリムシ(P.includens)、ツマジロクサヨトウ(S.frugiperda)及びサトウキビメイガ(D.saccharalis)からなる群から選択されるLepidopteran害虫に対して試験を行う際に、毒性の生物活性を示す、
前記Lepidopteran毒性タンパク質。
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